Real-time PCR assay for detection of Environmental DNA from <i>Chironex yamaguchii</i>

Abstract

e box jelly sh Chironex yamaguchii is a highly venomous cubomedusa, also known as "habu-kurage" in Japan. It is a signi cant threat to marine activities in Okinawa in southern Japan. Annually, envenomation of the species causes nearly 100 to 200 injuries in the Okinawa Prefecture. e present study developed a species-speci c real-time PCR assay to analyze environmental DNA (eDNA) for rapid detection of jelly sh in water samples. Primers and probe were designed based on the sequences of the mitochondrial cytochrome c oxidase subunit I region using tissue samples of nine specimens obtained from the Okinawa Prefecture. e speci city of the realtime PCR assay was con rmed by amplifying the DNA samples of 10 di erent box jelly sh species. A practical assay for eDNA detection was successful on water samples obtained from the natural habitat of C. yamaguchii. In addition, the degradation rate of eDNA in the tank experiment was 65% of the initial concentration at 1 h, 22% at 12 h, and 3% at 48 h.